小冠花高效体细胞胚胎发生与植株再生的研究

A study on efficcient somatic embryogenesis and plant regeneration in crownvetch

目的建立小冠花稳定高效体细胞胚胎植株再生体系。方法采用植物离体组织培养技术,通过优化外植体,基本培养基及其激素组合,提高体细胞胚发生和植株再生频率。结果子叶外植体在附加1.0mg/L 2,4-D,1.0mg/L NAA,0.5mg/L KT和100mg/L脯氨酸的MSB培养基上35d100%形成体细胞胚,产生体细胞胚数量达625个/g,在MS+KT 1.0mg/L+IBA 0.1mg/L+脯氨酸300 mg/L培养基上转换成完整植株的平均数为21棵/g。再生植株移栽成活率为80%,其形态特征和生长习性未见变异。外源激素是体细胞胚胎诱导所必需的,2,4-D是其中最关键的一种。结论建立了小冠花高频稳定高效体细胞胚胎发生和植株再生体系。

Aim To estabilish system of efficient plant regeneration via somatic embryogenesis in crownvetch （ Coronilla varia L. ）. Methods Optimization of explant types, basal media, and hormone combination to increase frequency somatic embryogenesis and plant regeneration by using plant tissue culture technology. Results High frequency （ 100% ） of somatic embryos with an average of 625 per gram fresh weight of embryogenic callus has been obtained from cotyledon-derived calli on modified MS medium containing 1 mg/L 2,4-D, 1 mg/L NAA, 0. 5 mg/L K and 100 mg/L proline within 35 d. Transfer of somatic embryos to MS medium supplemented with 1.0 mg/L KT, 0. 1 mg/L IBA, 300 mg/L proline and 1% activated charcoal resulted in complete plant regeneration at high frequency（ 100% ） with an average of 21 plantlets per gram fresh weight of calli in 5 weeks. Phytohormone was necessary to induced callus and somatic embryogenesis, and 2,4-D play a key role in embryos induction, while KT, NAA combined with 2,4-D had an important improving effect. The rooted plantlets were successfully transferred to soil with 80% survivor and the plants showed normal morphological and growth characteristics. Conclusion High efficient plant regeneration via somatic embryogenesis was achieved in crownvetch （ Coronilla varia L） .